Preparation of pMPMT5-AID+AIDT7 construct

Michał K.:

1. Digest of DNA fragment (AIDT7) with HindIII and SalI (2x Tango buffer).
2. Digest of pMPM-T5 + AID with HindIII and SalI(2x Tango buffer), dephosphorylation (CIAP).
3. Clean-up of reaction products.
4. Ligation of purified DNA (pMPM-T5 + AID with AID-T7 RNA-polymerase translation fusion).
5. Transformation of E.coli TOP10 with ligation product.
6. Plating transformants on LB+tetracycline.

Preparation of pMPMT5+T7 construct

Piotr:

1. Inoculation some separate transformant colonies (liquid LB + tetracycline).